Transfection deals with the introduction of foreign DNA into eukaryotic cells. In the lab, this is often done as an instance of transient transfection, in which the transfected gene is expressed only transiently. The gene is only expressed in the cell to which it was originally inserted and only for a brief period of time. This method is commonly used to test how various genetic modifications affect the functioning of particular genes. Find here protocols for transient transfection.
Transient Transfection and Luciferase assay Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay.
Transient Transfection Into 293T Cells Protocol Transient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) proteins. 293 is a human renal epithelial cell line which is transformed by adenovirus E1A gene product. 293T is a derivative which also express SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin and early promoter region. They (both) have the unusual property of being highly transfectable.
Transient Transfection: Viral Production This transient transfection protocol for the ecotropic/amphotropic producer line includes instructions for titering and concentration of viral stocks. This procedure gives consistently higher transfection efficiency.
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